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| Status |
Public on Aug 15, 2006 |
| Title |
Identifying targets of FLC at 27oC |
| Organism |
Arabidopsis thaliana |
| Experiment type |
Expression profiling by array
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| Summary |
FLOWERING LOCUS C (FLC) is a MADS box transcription factor that plays a well characterised role in repressing the vegetative to floral transition of Arabidopsis thaliana. FLC has also been shown to affect the Arabidopsis circadian clock, with mutant seedlings showing short circadian periods. In a previous study, we identified the temperature-dependent circadian period QTL PerCv5b near the FLC locus on the top arm of Chromosome 5. PerCv5b caused a significant period effect at 27oC but not at 12oC or 22oC. Temperature-dependent circadian period phenotypes and a known polymorphism in the Ler allele made FLC a strong candidate gene for PerCv5b. The period effect of FLC was enhanced by combination with alleles of FRIGIDA (FRI), a gene shown to up-regulate FLC's expression.
We were interested in identifying how FLC affects the circadian clock, so we decided to identify its target genes. Greatest phenotypic difference was observed between fri; flc and FRI; FLC genotype seedlings at 27oC, so expression was compared between these lines (previously described in Michaels and Amasino1999 and 2001) on the Affymetrix ATH1 microarray. Seedlings were grown on media (MS 1.5% Agar containing 3% Sucrose) for 6 days under constant cool white fluorescent light (55-60 micro EINSTEINS) at 22oC then entrained for 4 days under (12h , 12h) light , dark cycles at 22oC. At dawn on the fourth day of entrainment they were transferred to constant light (25-30 micro EINSTEINS) at 27oC. Four samples were taken at 6 hour intervals starting 24h after the transfer to continuous conditions at the times 24h, 30h, 36h and 42h.
Equal amounts of total RNA were pooled from the three time points to produce one sample per genotype. The pooling strategy was employed to reduce the effect of circadian regulation on genes expression. This was particularly important in our case as some interesting genes would likely be regulated by the circadian clock and may only show expression differences at particular phases that could easily be missed if using just one time point.
Experimenter name = Kieron Edwards
Experimenter phone = 0131 651 3326
Experimenter fax = 0131 650 5392
Experimenter department = Institute of Molecular Plant Sciences
Experimenter institute = University of Edinbugh
Experimenter address = Kings Buildings
Experimenter address = Mayfield Road
Experimenter address = Edinburgh
Experimenter zip/postal_code = EH9 3JH
Experimenter country = UK
Keywords: genetic_modification_design
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| Overall design |
4 samples were used in this experiment
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| |
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| Contributor(s) |
Edwards K, Millar A, Townsend H, Emmerson Z, Schildknecht B |
| Citation(s) |
16473970 |
| |
| Submission date |
Aug 15, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Nottingham Arabidopsis Stock Centre (NASC) |
| E-mail(s) |
[email protected]
|
| Phone |
+44 (0)115 951 3237
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| Fax |
+44 (0)115 951 3297
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| URL |
http://arabidopsis.info/
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| Organization name |
Nottingham Arabidopsis Stock Centre (NASC)
|
| Department |
School of Biosciences, University of Nottingham
|
| Street address |
Sutton Bonington Campus
|
| City |
Loughborough |
| ZIP/Postal code |
LE12 5RD |
| Country |
United Kingdom |
| |
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| Platforms (1) |
| GPL198 |
[ATH1-121501] Affymetrix Arabidopsis ATH1 Genome Array |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA96007 |
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